The Impact Of Tyrosine Kinase Inhibitor On The T Lymphocyte

Objective:To observe the effect of tyrosine kinase inhibitor(TKI)such as sunitinib,icotinib and erlotinib on T lymphocyte and to explore the possibility of the combination use of immunotherapy and tyrosine kinase inhibitor.Methods:T lymphocytes were selected from peripheral blood mononuclear cells by immunomagnetic cell sorting.Then the T lymphocytes were stimulated by Phorbol-12-myristate-13-acetate(PMA)and increasing dose of sunitinib,icotinib and erlotinib was added to the experimental group,respectively.After culturing 48-72 hours the proliferation rate of the T lymphocytes was analyzed by MTT.The subgroups of T lymphocyte,the apoptosis and cell cycle distribution were detected by flow cytometry(FCM).Supernatants of the cultured systems were collected to detect the levels of IFN-?,IL-2,IL-4,IL-10 and TGF-? by ELISA.Also the activity of cytotoxic lymphocyte(CTL)was detected by lactate dehydrogenase(LDH)release assay.Results:1.The effect of TKI to the proliferation of T lymphocyte:we compared the inhibitory rate between the control group and experiment group,and we found that TKI can inhibit the proliferation of T lymphocyte.2.The effect to the apoptosis of T lymphocytes:after cultured 48 hours we analyzed the apoptosis rate of the T lymphocytes by Annexin-V-PI method and we found that there was no significant differences between control group and experiment groups(P>0.05).3.The effect to the cell cycle:sunitinib,icotinib and erlotinib caused G0/G1 arrest in activated T cells.4.The effect to the subgroup of T lymphocytes:all of the three TKIs had no effect on the ration of CD4+ and CD8+T lymphocyte(P>0.05).Meanwhile the results showed that sunitinib can reduce percentage of Treg cells(P<0.05),while icotinib and erlotinib reduced percentage of Treg cells only in a high concentration(P<0.05).5.The changing of proinflammatory cytokines in activated T cells:sunitinib reduced the production of IFN-??IL-2?TGF-?,and lightly reduced IL-10,while there was no effect on the production of IL-4.Icotinib inhibited the production of TGF-?,but there was no effect on the production of IFN-??IL-2?IL-4?IL-10.Erlotinib reduced the production of IFN-??IL-2?TGF-?,while there was no effect on the production of IL-4?IL-10.6.The changing of the cytotoxicity of T lymphocyte on tumor cells:LDH essay showed that the cytotoxicity of T lymphocytes on tumor cells was reduced after cultured with TKIs..Conclusion:TKI can inhibit the.proliferation of T lymphocyte and reduce the production of cytokines by T lymphocyte,also they can decrease the percentage of Treg.The G0/G1 phase arrest in activated T cells may be one of the mechanisms for the inhibition of T lymphocyte proliferation.TKI reduced the cytotoxicity of T lymphocytes on tumor cells,multitarget TKI had greater effect than single target TKI.We should consider the influence on immune system when using TKI,maybe icotinib is a better choice for combining with immunotherapy.